WO2015118030A2 - Antibody-drug conjugates and immunotoxins - Google Patents
Antibody-drug conjugates and immunotoxins Download PDFInfo
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- WO2015118030A2 WO2015118030A2 PCT/EP2015/052341 EP2015052341W WO2015118030A2 WO 2015118030 A2 WO2015118030 A2 WO 2015118030A2 EP 2015052341 W EP2015052341 W EP 2015052341W WO 2015118030 A2 WO2015118030 A2 WO 2015118030A2
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- KNVPTEBUNOFRAO-DZGJXGEKSA-N CCCN(C(C)(C[C@H](c1nc(C(N[C@H](C[C@@H](C(NN)=O)c2c3)Cc2ccc3O)=O)c[s]1)OCCC)C(C)C)C(C[C@H](CC)COC([C@@H]1N(C)CCCC1)N)=O Chemical compound CCCN(C(C)(C[C@H](c1nc(C(N[C@H](C[C@@H](C(NN)=O)c2c3)Cc2ccc3O)=O)c[s]1)OCCC)C(C)C)C(C[C@H](CC)COC([C@@H]1N(C)CCCC1)N)=O KNVPTEBUNOFRAO-DZGJXGEKSA-N 0.000 description 1
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- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
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Definitions
- the present invention relates to antibody-drug conjugates (ADCs) and Immunotoxins that target Fibroblast Activating Protein a (FAP) , and to their use in medicine, e.g. in the treatment of certain cancers.
- ADCs antibody-drug conjugates
- FAP Fibroblast Activating Protein a
- Malignant epithelial tumors are the main cancer-related cause of human death. These solid tumors frequently exhibit significant stromal reactions such as the so-called “desmoplastic stroma” or “reactive stroma”, which represents 20-60% of total tumor mass and is characterized by the existence of large numbers of stromal cells and dense extracellular matrix (ECM) . Recent studies have indicated the tumor-promoting roles of stromal cells, as exemplified by vascular cells, immune cells, fibroblasts, myofibroblasts,
- CAFs cancer-associated fibroblasts
- CAFs facilitate the spreading and infiltration of tumor cells in distant organs, thus contributing to formation of metastases.
- the relevance of stromal cells to the failure of systemic drug delivery to tumors and to the development of drug resistance has also been indicated (7-11).
- MAb Monoclonal antibody
- chemotherapeutic molecules can be good candidates, since they are very potent and active in very small quantities .
- Immunoconjugates are made of a human, humanized or chimeric
- the Ab must be very selective to reach the antigen, whose expression must be restricted in normal cells.
- the Ab also must be internalized efficiently into the cancerous cells.
- the cytotoxic agent selected as the effector moiety must kill cells only after internalization and release into the cell cytoplasm.
- the most commonly used payloads in ADCs are DNA-harming drugs such as calicheamicins , duocarmicins , or microtubule-targeting compounds like auristatins and maitansinoids .
- Ab-cytotoxic linkers are designed to be stable systemically and release the cytotoxic within the target cells.
- TAAs are frequently cell membrane proteins that are overexpressed in diseased tissues or at least expressed sufficiently to facilitate the internalization-activated cytotoxicity.
- the antigen presents a restricted expression in normal tissues with a low or absent expression in vital organs.
- the tumor antigen must be recognized selectively and with high affinity by an Ab.
- fibroblast response is characterized by the induction of a cell surface protein, Fibroblast Activating Protein a (FAPa), a serine protease of 95kDa whose expression is highly restricted to developing organs, wound-healing and tissue remodeling .
- FAPa Fibroblast Activating Protein a
- FAP expression has been recently found in Pancreas tumor cells as well as tumor-associated stromal fibroblasts. FAP expression was correlated with shorter patient survival and worse prognosis, suggesting a possible FAP-based autocrine/paracrine loop in this type of tumor (32) .
- the present invention relates to anti-FAP antibodies, conjugates thereof and optimised payloads for use in antibody conjugate strategies.
- anti-FAP antibodies as described herein exhibit highly specific binding, and fast and efficient internalisation .
- the present inventors have found that the A chain of Nigrin b can be isolated and produced in bacterial host cells, yet retains in vitro Ribosome Inactivating activity in the absence of the Nigrin-b B- chain and, only once conjugated to an antibody, exhibits both the ability to translocate into cells and the resulting cytotoxic activity without Nigrin-b B-chain.
- the present invention provides a conjugate having the formula I:
- L is a linker
- D is a drug comprising a cytolysin or a Nigrin-b A-chain; and p is 1 to 10.
- CDRH1 SEQ ID NO: 7 or a variant thereof having up to 1 or 2 amino acid substitutions compared with the sequence of SEQ ID NO: 7
- CDRH2 SEQ ID NO: 8 or a variant thereof having up to 1 or 2 amino acid substitutions compared with the sequence of SEQ ID NO: 8
- CDRH3 SEQ ID NO: 9 or a variant thereof having up to 1 or 2 amino acid substitutions compared with the sequence of SEQ ID NO: 9
- CDRL1 SEQ ID NO: 10 or a variant thereof having up to 1 or 2 amino acid substitutions compared with the sequence of SEQ ID NO: 10
- CDRL2 SEQ ID NO: 11 or a variant thereof having up to 1 or 2 amino acid substitutions compared with the sequence of SEQ ID NO: 11;
- CDRHl-3 comprise the amino acid sequences of SEQ ID NOS : 7-9, respectively and CDRLl-3 comprise the amino acid sequences of SEQ ID NOS: 10-12, respectively.
- A comprises a heavy chain variable region (VH) comprising an amino acid sequence having at least 90%, 95% or 99% sequence identity with the full-length sequence of SEQ ID NO: 5.
- VH heavy chain variable region
- A comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 5.
- A comprises a light chain variable region (VL) comprising an amino acid sequence having at least 90%, 95% or 99% sequence identity with the full-length sequence of SEQ ID NO: 6.
- VL light chain variable region
- A may comprise a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 6.
- A comprises a heavy chain comprising an amino acid sequence having at least 90%, 95% or 99% sequence identity with the full-length sequence of SEQ ID NO: 3.
- A may comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 3.
- A comprises a light chain comprising an amino acid sequence having at least 90%, 95% or 99% sequence identity with the full-length sequence of SEQ ID NO: 4.
- A may comprise a light chain comprising the amino acid sequence of SEQ ID NO: 4.
- A may be a competitively binding anti-FAP antibody that is structurally different from the anti-FAP antibody molecules exemplified herein.
- A may be an anti-FAP antibody molecule that competes with the anti-FAP IgGl antibody identified herein as "hu36" for binding to immobilized recombinant human FAP .
- hu36 has the heavy chain amino acid sequence of SEQ ID NO: 3 and the light chain amino acid sequence of SEQ ID NO: 4.
- the anti-FAP antibody may, in some case, bind to the same epitope as hu36.
- D may be a cytolysin.
- the cytolysin may, in some cases, be a compound disclosed in WO 2008/138561 Al, the entire contents of which is expressly incorporated herein by reference (compounds disclosed therein are also referred to as Tubulysine derivatives) .
- the cytolysin may be synthesised as described in WO 2008/138561.
- the cytolysin may be as defined in Formula I or Formula IV of WO 2008/138561 Al .
- the cytolysin may be of formula IV:
- R 2 (i) is directly or indirectly attached to linker L or (ii) is H or is C1-C4 alkyl;
- R 6 is Ci-C 6 alkyl
- R 7 is Ci-C 6 alkyl, CH 2 OR 19 or CH2OCOR 20 , wherein R 19 is alkyl, R 20 is C 2 - C6-alkenyl, phenyl, or CH 2 -phenyl;
- R 9 is Ci-C 6 alkyl
- R 10 is H, OH, O-alkyl or O-acetyl
- f 1 or 2;
- R 11 has the following structure:
- R 21 is H, OH, halogen, NH 2 , alkyloxy, phenyl, alkyl amino or dialkyl amino ;
- R 16 is H or a Ci-C6-alkyl group
- R 17 (i) is directly or indirectly attached to linker L or (ii) is C0 2 H, C0 2 R 18 , CONHNH2, OH, NH 2 , SH or a optionally substituted alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl group, wherein R 18 is an optionally substituted alkyl, heteroalkyl or hetercycloalkyl group; and
- C1-C6 alkyl substituted with unsubstituted C1-C6 alkyl, C2C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C3-C10 cycloalkyl, C2-C9 heterocycloalkyl, C6-C10 aryl, C1-C9 heteroaryl, C7-C12 aralkyl or C2-C11 heteroaralkyl groups .
- R 2 is a bond to linker L.
- R 17 is C(0)X, CONHNHX, OX, NHX or SX, wherein X is a bond to linker L.
- linker L may further comprise a spacer.
- the spacer has a chain length of 2 to 30 atoms.
- the spacer comprises or consists of an alkylene (i.e. divalent alkyl) or heteroalkylene (i.e. divalent heteroalkyl) group. In some cases the spacer comprises or consists of an alkylene or oxyalkylene group.
- the spacer comprises or consists of a group -(CH 2 ) n - or
- the spacer comprises or consists of a group
- n may be 1 to 15, 1 to 10, 1 to 6, or 2 to 5.
- n may be 3 or 4.
- the spacer comprises between one and six ethylene glycol units, e.g. a triethylene glycol.
- the spacer is attached to group R 17 via a -C(0)X bridging group, wherein X is a bond to R 17 .
- R 17 is CONHNHX and the spacer is attached to group R 17 via a -C(0)X bridging group, wherein X represents the bond between the spacer and R 17 .
- D comprises a cytolysin having the following
- L comprises an attachment group for attachment to A and protease cleavable portion.
- L may comprise a valine-citrulline unit.
- L may comprise
- the double bond of the maleimide is reacted with a thiol group of a cysteine residue of the antibody A to form a sulphur-carbon bond in order to effect linkage of the linker L to the antibody A.
- -L-D has a structure selected from the group
- -L-D may have the following structure:
- p may, in some cases, lie in the range 1 to 5, e.g. 1 to 4, or 1 to 3. In particular cases p may be 1 or 2. In particular, cases p may be 3 or 4.
- D may be a Nigrin-b A-chain.
- the Nigrin-b A-chain is in the absence of a Nigrin-b B-chain.
- the Nigrin-b A-chain may comprise or consist of the sequence of SEQ ID NO: 13.
- L may simply be a disulphide bond between a sulphur atom on A and a sulphur atom on D.
- L may comprise or consist of a disulphide bond.
- the present invention provides a conjugate as defined in accordance with the first aspect of the invention for use in medicine.
- the present invention provides a conjugate as defined in accordance with the first aspect of the invention for use in a method of treatment of a tumor in a mammalian subject.
- chemotherapeutic agent or an anti-angiogenic agent or an
- the one or more other antitumor drugs comprise Gemcitabine, Abraxane bevacizumab,
- itraconazole carboxyamidotriazole , an anti-PD-1 molecule or an anti-PD-Ll molecule (for example, nivolumab or pembrolizumab) .
- the present invention provides use of a cytolysin in the preparation of an antibody-drug conjugate, wherein the antibody is an FAP-specific antibody, e.g., an FAP-specific antibody in accordance with the eighth aspect of the invention.
- the use may be of a cytolysin in the preparation of an antibody-drug conjugate as defined in accordance with the first aspect of the invention .
- the present invention provides a conjugate of the first aspect of the invention for use in the treatment of an inflammatory condition (e.g. rheumatoid arthritis) .
- an inflammatory condition e.g. rheumatoid arthritis
- the present invention provides a method of treating an inflammatory condition (e.g. rheumatoid arthritis) in a mammalian subject, comprising administering a therapeutically effective amount of a conjugate of the first aspect of the invention to the subject in need thereof.
- an inflammatory condition e.g. rheumatoid arthritis
- the present invention provides an isolated Nigrin-b A-chain in the absence of the Nigrin-b B-chain.
- the amino acid sequence of the Nigrin-b A- chain may comprise or consist of the sequence of SEQ ID NO: 13.
- the present invention provides use of an isolated Nigrin-b A-chain in accordance with the eighth aspect of the invention in the preparation of an immunotoxin.
- the immunotoxin comprises a monoclonal antibody conjugated and/or bound to said isolated Nigrin-b A-chain (in the absence of the Nigrin-b B- chain) .
- the immunotoxin comprises an antibody, such as a monoclonal antibody, e.g. a human monoclonal antibody, that selectively binds FAP .
- the immunotoxin comprises an antibody in accordance with the tenth aspect of the invention.
- the present invention provides a monoclonal antibody, e.g. a human monoclonal antibody, that selectively binds FAP and which comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain comprising the amino acid sequence of SEQ ID NO: 4.
- the present invention provides use of a
- the present invention provides a host cell comprising a vector comprising a polynucleotide that encodes at least one polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NOS : 1-6 and 13.
- the polynucleotide may comprise the nucleic acid sequence of SEQ ID NO:
- the process may further comprise a step (c) of purifying and/or isolating the conjugate.
- an appropriate residue e.g. a cysteine amino acid
- step (a) may comprise reacting the antibody with 4- succynimidyloxycarbonyl-a-methyl- ⁇ - ( 2-pyridyl-dithio ) toluene (SMPT) , N-succynimidyl 3— (2-pyridyl-dithiopropionate) (SPDP) or methyl 4- mercaptobutyrimidate .
- SMPT 2-pyridyl-dithio toluene
- SPDP N-succynimidyl 3— (2-pyridyl-dithiopropionate
- the present invention provides a process for the production of a conjugate in accordance with the first aspect of the invention, comprising:
- the process may comprise purifying and/or isolating the conjugate .
- Figure 1 shows characterization of humanized scFv hu33 and hu36.
- B) Flow cytometry analysis of binding of hu36 (humanized) and mo36 (chimeric) to HT1080-huFAP cells. Bound antibodies were detected with an anti-His-tag antibody (n 2) .
- Figure 2 shows A) ELISA of anti-FAP mo36-IgGl (circles) and hu36- IgGl (squares) for binding to recombinant human FAP (rhFAP) or control protein (BSA) (triangles and inverted triangles, respectively) . 50 ng protein were coated per well. Bound antibodies were detected with HRP-conj ugated anti-human IgG-Fc. B) Flow cytometry analysis of anti-FAP mo36-IgGl (triangles and stars) and hu36-IgGl (squares and circles) for binding to HT1080-FAP. Bound proteins were detected with a PE-labeled anti-hu IgG-Fc antibody.
- Figure 3 shows flow cytometry analysis of binding of hu36-IgGl to stably transfected HT1080 to express A) human FAP (HTl 080-huFAP) and B) mouse FAP (HTl 080-moFAP) . Bound antibodies were detected with a PE-labeled anti-human Fc antibody.
- Figure 5 shows analysis of internalization of hu36-IgGl by
- Figure 6 shows MALDI-Tof profile of recombinant nigrin-b A-chain. Observed mass (Da) : 28546.55; Expected mass (Da) : 28546.09; Mass deviation: 0.5; Mass Accuracy: 16ppm.
- Figure 7 shows ribosome inactivating protein (RIP) activity of recombinant Nigrin-b A-chain (recNgA) tested in rabbit reticulocyte cell-free lysates (RRL) versus native (WT) Nigrin-b. (3a, 3b, 6c, 9c) represent different formulations of recNgA.
- RIP ribosome inactivating protein
- Figure 8 shows cytotoxicity of recNgA tested on HT1080-FAP cell line through crystal violet viability assay (native Nigrin - diamonds; recombinant Nigrin-b A-chain - squares) .
- Figure 9 shows RIP activity of anti-FAP hu36-IgGl-recNgbA immunotoxin conjugates (HSP131-001 ; crosses) in an RRL assay compared to native (WT) nigrin (triangles) and recombinant Nigrin-b A-chain (recNgA; squares) .
- Figure 10 shows cytotoxic activity of anti-FAP hu36-IgGl-recNgbA immunotoxin conjugates (HSP131-001 ; triangles), unconjugated
- Figure 11 shows the general antibody conjugate structure for a cytolysin-conjugated antibody via a vcPABA linker. Attachment of the cytolysin may be via Ri or R4 (identified by arrows) .
- Figure 12 shows immunodetection of anti-FAP hu36 tumour sections of patient-derived xenograft (PDX) mice (pancreatic tumour) . Specific Dose- and Time- dependent staining of stroma is observed in
- Figure 13 shows animal weight monitored after treatment with anti- FAP:recNgA immunotoxin at different doses (2.5, 1, 0.5, 0.25, 0.1 mg/kg) administrated once a week. Significant weight loss and toxicity was observed in Group 1 and 2 (2.5 and 1 mg/kg,
- 0.5mg/kg was the highest tolerated dose when applied as single agent .
- Figure 14 shows (A) Relative Body weight and (B) Tumor volume measured from patient-derived xenograft mice (PAXF 736) untreated (Vehicle; 1 Oml/kg/day; once a week), treated with Gemcitabine (GEM; 150 mg/kg; once a week), or antiFAP : recNgA immunotoxin (OMTX505; 0.5/0.25mg/kg; once a week), or both (OMTX505
- Figure 15 shows ELISA and FACS analysis of ADC471 binding to FAP target.
- B & (C) : FACS analysis of binding on HT1080-huFAP, HT1080-wt and HEK293 cells of HPS131-143-1 (ADC-471; DAR 4), HPS131-124-1 (ADC-467; DAR 1.2) and HPS131-124-3 (ADC-471; DAR 3.48) ADCs.
- EC50 values are indicated for this latter (B) .
- FIG. 16 shows Time-lapse immunofluorecence analysis
- anti-FAP hu36 cytolysin ADC (ADC-471; HPS131-124-3) on living HT1080-FAP cells. Left panel: Incubation with naked anti-hu/moFAP hu36 (FITC-AB; green); Right panel:
- Figure 17 shows in vitro cytotoxic effect of anti-hu/moFAP hu36: cytolysin ADCs on (A) HT1080-wt and (B) FAP(+) cells.
- Figure 18 shows tumor growth inhibition effect of anti-hu/moFAP hu36: cytolysin ADC candidates.
- A ADC471 versus ADC551;
- B ADC471 and ADC553 (OMTX705-553) versus ADC558 (OMTX705-558 ) .
- Vehicle and GEM Gemcitabine: negative and positive control groups.
- Fibroblast activation protein As used herein "Fibroblast activation protein", "fibroblast
- FAP activating protein
- FAPa FAP activating protein activating protein
- the FAP may be an FAP of any mammalian species.
- FAP is human FAP (also known as Seprase, 170 kDa melanoma membrane-bound gelatinase, fibroblast activation protein alpha or integral membrane serine protease) , the amino acid sequence of which is disclosed at UniProt accession No. Q12884 (Version 140, dated 11 December 2013)
- a molecule that binds FAP may bind to a region of the extracellular domain of FAP.
- the extracellular domain of human FAP comprises residues 26-760 of the full-length human FAP protein.
- FAP is murine FAP (also known as fibroblast activation protein alpha or integral membrane serine protease) , the amino acid sequence of which is disclosed at UniProt accession No. P97321
- extracellular domain of murine FAP comprises residues 26-761 of the full-length murine FAP protein.
- conjugate includes the resultant structure formed by linking molecules and specifically includes antibody-drug conjugates (ADCs) and immunotoxins (ITs) .
- ADCs antibody-drug conjugates
- ITs immunotoxins
- the terms selectively binds and selective binding refer to binding of an antibody, or binding fragment thereof, to a predetermined molecule (e.g. an antigen) in a specific manner.
- the antibody, or binding fragment thereof may bind to FAP, e.g. an extracellular portion thereof, with an affinity of at least about lxl0 7 M _1 , and may bind to the predetermined molecule with an affinity that is at least two-fold greater (e.g. five-fold or ten-fold greater) than its affinity for binding to a molecule other than the predetermined molecule.
- antibody or “antibody molecule” includes any immunoglobulin whether natural or partly or wholly synthetically produced.
- antibody or “antibody molecule” includes monoclonal
- Antibodies may be intact or fragments derived from full antibodies (see below) .
- Antibodies may be human antibodies, humanised antibodies or antibodies of non-human origin.
- “Monoclonal antibodies” are homogeneous, highly specific antibody populations directed against a single antigenic site or "determinant" of the target molecule.
- “Polyclonal antibodies” include heterogeneous antibody populations that are directed against different antigenic determinants of the target molecule.
- antigen or
- antigena refers to blood serum containing antibodies obtained from immunized animals.
- binding fragments are (i) the Fab fragment consisting of VL, V H , CL and CHI domains; (ii) the Fd fragment consisting of the V H and CHI domains;
- the term “selectively binds” may be used herein to refer to the situation in which one member of a specific binding pair will not show any significant binding to molecules other than its specific binding partner (s) .
- the term is also applicable where e.g. an antigen-binding site is specific for a particular epitope that is carried by a number of antigens, in which case the specific binding member carrying the antigen-binding site will be able to bind to the various antigens carrying the epitope.
- the antibody may be a fully human antibody.
- the conjugate of the invention may administered with, or for administration with, (whether simultaneously, sequentially or separately) one or more other antitumor drugs, including, but not limited to, a cytotoxic chemotherapeutic agent or an anti-angiogenic agent or an immunotherapeutic agent.
- one or more other antitumor drugs including, but not limited to, a cytotoxic chemotherapeutic agent or an anti-angiogenic agent or an immunotherapeutic agent.
- Cytotoxic chemotherapeutic agents are well known in the art and include anti-cancer agents such as:
- Alkylating agents including nitrogen mustards such as
- mechlorethamine HN2
- cyclophosphamide ifosfamide, melphalan (L- sarcolysin) and chlorambucil
- 10 ethylenimines and methylmelamines such as hexamethylmelamine , thiotepa
- alkyl sulphonates such as busulfan
- nitrosoureas such as carmustine (BCNU) , lomustine (CCNLJ) , semustine (methyl-CCN-U) and streptozoein ( streptozotocin)
- triazenes such as decarbazine (DTIC;
- Antimetabolites including folic acid analogues such as methotrexate (amethopterin) ; pyrimidine analogues such as fluorouracil (5- fluorouracil ; 5-FU) , floxuridine ( fluorodeoxyuridine ; FUdR) and cytarabine (cytosine arabinoside) ; and purine analogues and related inhibitors such as mercaptopurine ( 6-mercaptopurine ; 6-MP) ,
- thioguanine 6-thioguanine ; TG
- pentostatin (2'- deoxycofonnycin)
- Natural Products including vinca alkaloids such as vinblastine (VLB) and vincristine; epipodophyllotoxins such as etoposide and tenyposide; antibiotics such as dactinomycin
- doxorubicin doxorubicin
- bleomycin bleomycin
- plicamycin mithramycin
- mitomycin Q enzymes such as L-asparaginase ; and biological response modifiers such as interferon alphenomes .
- Miscellaneous agents including platinum coordination complexes such as cisplatin
- hydrazine derivative such as procarbazine (N- methylhydrazine, MIH) ; and adrenocortical suppressant such as mitotane (o, p'-DDD) and aminoglutethimide ; taxol and analogues/derivatives; and hormone agonists/antagonists such as flutamide and tamoxifen.
- a further preferred cytotoxic agent is Gemcitabine (Gerazar®) .
- a further preferred cytotoxic agent is Paclitaxel bound to human serum albumin
- Anti-angiogenic agents are well known in the art and include anticancer agents such as bevacizumab, itraconazole, and
- Immunotherapeutic agents include, for example, anti-programmed cell death protein 1 (PD-1) antibodies and anti-programmed death-ligand 1 (PD-L1) antibodies, including
- Nivolumab (MDX1106) and Pembrolizumab (MK-3475) .
- the conjugates of the present invention may be comprised in
- compositions with a pharmaceutically acceptable excipient .
- a pharmaceutically acceptable excipient may be a compound or a combination of compounds entering into a pharmaceutical composition which does not provoke secondary reactions and which allows, for example, facilitation of the administration of the conjugate, an increase in its lifespan and/or in its efficacy in the body or an increase in its solubility in solution.
- These pharmaceutically acceptable vehicles are well known and will be adapted by the person skilled in the art as a function of the mode of administration of the conjugate.
- conjugates of the present invention may be provided in a lyophilised form for reconstitution prior to
- lyophilised conjugates may be reconstituted in sterile water and mixed with saline prior to
- compositions may comprise, in addition to the conjugate, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the conjugate.
- a pharmaceutically acceptable excipient such materials should be non-toxic and should not interfere with the efficacy of the conjugate.
- carrier or other material will depend on the route of
- administration which may be by bolus, infusion, injection or any other suitable route, as discussed below.
- composition comprising the conjugate may be in the form of a parenterally acceptable aqueous solution which is pyrogen- free and has suitable pH, isotonicity and stability.
- a parenterally acceptable aqueous solution which is pyrogen- free and has suitable pH, isotonicity and stability.
- isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection.
- Preservatives, stabilizers, buffers, antioxidants and/or other additives may be employed as required including buffers such as phosphate, citrate and other organic acids; antioxidants, such as ascorbic acid and methionine; preservatives (such as
- octadecyldimethylbenzyl ammonium chloride hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol ; 3'-pentanol; and m-cresol); low molecular weight polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers, such as
- the subject may be a human, a companion animal (e.g. a dog or cat), a laboratory animal (e.g. a mouse, rat, rabbit, pig or non-human primate), a domestic or farm animal (e.g. a pig, cow, horse or sheep) .
- the subject is a human.
- the subject may be a human diagnosed with or classified as being at risk of developing a cancer, e.g., an epithelial tumor.
- the subject may be a laboratory animal, e.g., a mouse model of a cancer.
- the subject may be a mammal (e.g. a human) that has been diagnosed with or classified as being at risk of developing an inflammatory condition, such as rheumatoid arthritis (RA) .
- the subject may be a human having RA.
- the anti-FAP conjugates described herein find use in the treatment of a tumor in a mammalian subject.
- the tumor may be a solid tumor.
- the tumor may be a pancreatic cancer, breast cancer, melanoma, lung cancer, head & neck cancer, ovarian cancer, bladder cancer or colon cancer.
- Anti-FAP scFvs selected by phage display from an immunized FAP _/_ knock-out mouse have been described previously (28) .
- Two scFvs, "M036” and “M033", cross-reactive for human and murine FAP (28) were converted into full-length IgG for subsequent characterisation studies and for generation of immunotoxins and ADCs .
- These scFv (scFv33 and scFv36) were used to generate chimeric antibodies, fusing heavy and light chain constant domains to VH and VL,
- scFvs were bacterially produced in E. coli TGI and purified from the periplasmic extracts of 1L cultures by IMAC . Both humanized antibodies (scFv hu33 and hu36) were purified in soluble form with yields of approximately 0.6mg/L culture. In SDS-PAGE the proteins migrated with the expected size of approximately 30kDa ( Figure 1A) . Purity was estimated to be >90%. In flow cytometry experiments using HT1080 cells expressing human FAP (stable transfectants ) , a similar binding was observed for scFv hu36 and mo36 scFv, which was also produced in bacteria (not shown) . EC50 values were in the low nanomolar range. Some differences were observed at higher
- Plasmids corresponding to full length IgGl antibodies were generated and transfected into CHO cells for production of antibodies in Lonza's CHO expressing system with yields of approximately 1 mg/L of cell culture (lab scale) .
- Antibodies were purified from cell culture supernatant by protein A chromatography. Purified proteins were characterized by SDS-PAGE and size exclusion chromatography.
- Bioactivity was analyzed by ELISA using recombinant FAP and
- VH domain is underlined; CDRH1-H3 are shown in bold and curved underlined .
- VL domain is underlined; CDRL1-L3 are shown in bold and curved underlined .
- hu36-IgGl-HC - without signal sequence
- ENIIH (SEQ ID NO: 7) hu36-CDRH2 :
- LASNLES SEQ ID NO: 11
- the antibody constructs were cloned in GS double vectors (pEE14.4) .
- the DNA plasmids were transformed, amplified, and transiently transfected into CHOK1SV cells for expression evaluation at a volume of 200 ml.
- the antibodies were transiently expressed in 5-10 L large scale cultures.
- Clarified culture supernatant was purified using one-step Protein A chromatography.
- Product quality analysis through SE-HPLC, SDS-PAGE and LAL was carried out using purified material at a concentration of 1 mg/ml, alongside an in-house human antibody as a control sample .
- the purified protein samples were filtered through a 0.2 ⁇ filter and analysed by SE-HPLC chromatograms .
- the antibodies were purified to >98.8%.
- the endotoxin levels were ⁇ 0.5 EU/mg.
- Purified proteins hu36-IgG and mu36-IgG were characterized by SDS- PAGE and size exclusion chromatography. Bioactivity was analyzed by ELISA, using recombinant FAP and detection of bound antibodies with HRP-conj ugated anti-human IgG antibodies. Cell binding was analyzed by flow cytometry, using HT1080-FAP cell line. Melting points were determined by dynamic light scattering using a zetasizer nano .
- the full-length IgGl purified antibodies were successfully produced at both lab scale and large scale, for the generation of
- Anti-FAP IgGl hu36 was administrated intraperitoneally to
- the enzymatic domain of Nigrin b was cloned and expressed in bacteria.
- Nigrin-b A-chain was synthetized taking into account codon
- Nigrin pET30b-3 and Nigrin pET33b-l (+/- His tag) for expression in two different E. coli strains, E. coli BLR ( DE3 ) and E. coli HMS174 (DE3) .
- E. coli BLR DE3
- E. coli HMS174 DE3
- Different culture media were used to check different expression conditions.
- Process purification was established using Capto Q chromatography and SP Sepharose High Performance. Purified recombinant Nigrin-b A-chain (recNgA) was formulated at 5mg/ml in PBS IX pH7.4, DTT 0.5 mM, glycerol 10%.
- Endotoxin levels were ⁇ lEU/mg of Nigrin and the purity >99% in monomeric form.
- the recombinant Nigrin-b A-chain has the following characteristics: Number of amino acids: 256
- Equilibration buffer A Glycine/NaOH 50 mM pH9.5.
- Elution buffer B Glycine/NaOH 50mM pH9.5, NaCl 1 M.
- AIM Auto Inducible Medium
- each cell pellet was initially resuspended in 80ml of extraction buffer per liter of culture, and 3 cycles of 7 minutes disintegration at 1100-110 Bar were performed after 30 minutes of incubation at 8°C under shaking. Then the extract underwent 60 minutes centrifugation at 15,900g, 8°C. The supernatant was the purification's starting material.
- Capto Q FPLC 160ml of extracted product from 81 culture were loaded into 160ml Capto Q and equilibrated using 4CV of equilibration buffer and washed with 15CV of equilibration buffer. Elution was carried in three steps: 15CV at 1.5mS/cm (7.6%B); 20CV at 23.8 mS/cm (18.9%B) ; 20CV 100%B.
- Dialysis was performed at the following conditions: 650ml of the product were dialyzed in 4x5Lbathsin in citric acid/NaOH 25 mM pH5.0, cut-off 6-8000Da. Dialysis factor -3500, ⁇ 24h.
- SP Sepharose HP 610ml of dialyzed pool of Capto Q in Citric acid 25 mM pH5.0 were loaded into 240ml SP Sepharose High Performance with 4CV of equilibration buffer and washed with 15CV of
- recNgA is stable at pH ranging from 5 to 9, and in presence or not of glycerol (from 10 to 45%) (data not shown) .
- the ribosome-inactivating protein (RIP) activity of recombinant Nigrin-b A-chain was tested in rabbit reticulocyte cell-free lysates: IC50 value obtained was similar to native nigrin-b and within 2.5 to 25 pM range (see Figure 7) .
- the A chain from Nigrin-b expressed as a recombinant protein in bacteria, maintains its enzymatic activity, supporting that glycosylation is not required for RIP activity of Nigrin-b A-chain.
- recNgA The cytotoxic activity of recNgA was tested on cell cultures through crystal violet-based viability assay.
- recNgA lacking the B chain to translocate within cells, presents a 100 to 1000 less toxic activity than native Nigrin-b, as shown in Figure 8.
- Native nigrin b showed an IC5o 3 ⁇ 4 2xlO _8 M (similar to previous published data; see 33)
- recNgA showed an IC 5 o ⁇ 2xlO- 6 M .
- Nigrin b presents higher RIP activity than Ricin in RRL assay, while it is much less toxic (30-10,000 time, approximately) in cells or in vivo (see IC50 and LD50 values in Table 3) .
- Nigrin b A chain generated for the first time in this present invention, only loses activity in cell cytotoxicity assay, while it was even increased in RRL assay with respect to native Nigrin b.
- Nigrin b A chain is 50 times more active than the Ricin A chain in RRL.
- Nigrin b A chain conjugates present higher cytotoxic activity (IC50 within pM range) than Ricin A chain conjugates (nM range) (data not shown) .
- Table 3 In vitro and in vivo activity data for Ricin and Nigr (native and A chain) .
- SPDP has already been used in the making of immunotoxins (ITs) containing nigrin b (38, 39) . Moreover SMPT protects the disulfide bond from attack by thiolate anions, improving in vivo stability of the linkage (40, 41) .
- DTT Dithiothreitol
- the recNgA sample was incubated in the presence of 4.8 mM DTT at room temperature for 30 min.
- the A280 of the aliquots was taken and the two most concentrated mixed.
- A280 was taken again.
- hetero-bifunctional reagents chemically inserted using hetero-bifunctional reagents, and several methods have been developed in order to generate hetero-conj ugates avoiding or reducing to a minimum the formation of homopolymers .
- the reagents used to introduce thiol groups react with amino groups, forming amide or amidine bonds. Amino groups are reactive, abundant and, in a limited way for most proteins,
- SPDP N-succynimidyl 3— (2-pyridyl-dithiopropionate)
- SMPT 4-succynimidyloxycarbonyl-a-methyl-a- (2-pyridyl- dithio) tolu
- SPDP and SMPT introduce hindered disulphide bond, while 2- iminothiolane -SH must be protected by reacting it with 5,5'- dithiobis-2-nitrobenzoic acid (Ellman' s reagent) .
- the reaction with Ellman' s reagent is also used for the quick measurement of protein sulphydryl groups (43, 44) .
- SMPT has a methyl group and a benzene ring attached to the carbon atom adjacent to disulphide bond that protects it from attack by thiolate anions, thus improving the in vivo stability of the linkage (40, 41) .
- IgG proteins can be modified with SMPT, which do not significantly affect the antigen binding property of the molecules in the following conditions, even if they change the charge of the protein in the reaction site.
- the immunotoxin is predominantly a mixture of antibody linked to one or two toxin molecules, with the presence of high molecular weight components (IgG linked to several RIP proteins) , as well as free and polymeric RIPs (dimeric in the case of recNgA) and free antibody.
- IgG linked to several RIP proteins high molecular weight components
- free and polymeric RIPs dimeric in the case of recNgA
- Anti-FAP hu36- IgGl -recNgA immunotoxin conjugates were produced and characterized as follows:
- Drug antibody ratio (DAR) : 1.8
- the RRL assay results show that the anti-FAP hu36-IgGl-recNgA conjugates (HPS131-001-1 ) presented similar IC50 values as native Nigrin-b or recNgA and were in the 3pM range, showing that antibody conjugation did not diminish the enzymatic activity of recNgA (see Figure 9) .
- the cell cytotoxicity results show that, on HT1080 wild-type cells, conjugated antibody HPS131-001-1 displays only slight toxicity (if any) and only at highest concentration, naked anti-FAP hu36-IgGl does not have any effect, and recNgA shows cytotoxic effect only at 10 "6 M and after 72h incubation (see Figure 10A) .
- HPS131-001-1 conjugated anti-FAP antibodies strongly reduce HT-1080-FAP cell viability in the picomolar concentration range, with IC50 values 5pM (see Figure 10B) .
- Immunotoxin anti-FAP recNgA has been tested in vivo in both cell- derived and patient-derived xenograft mouse models for pancreas cancer.
- a dose range study was first performed to define the maximum tolerated dose in normal mice and each of these models: doses from 5 to 0.1 mg/kg were administrated intraperitoneally once a week during 3 weeks, and animal weight was monitored every 2 days to detect possible weight loss due to toxic effect of the immunotoxin. Results are presented in Figure 13.
- Tubulysins are recently discovered natural compounds isolated from Myxobacteria, able to destabilize the tubulin skeleton, inducing apoptosis with a very high activity.
- tubulysins and their synthetic tetrapeptidic analogues are highly potent cell-killing agents (nM to pM activity) .
- Tubulysin A inhibits tubulin polymerization in vitro with an IC50 of 0.75-1 ⁇ , thus blocking the formation of mitotic spindles and inducing cell cycle arrest in G2/M phase.
- Tubulysins compete strongly with vinblastine through binding on the vinblastine binding site of tubulin. Furthermore they are stable in lysosome enriched cell fractions ⁇ 45-48) .
- Ri or R 4 - the Ri and R4 numbering system used in Figure 11 differs from the R group numbering system used, e.g., in the claims; it is intended that Ri of Figure 11 corresponds to R 2 in the claims and that R4 of Figure 11 corresponds to R 17 of the claims) .
- vcPABA valine-citrulline-PABC protease-cleavable linker
- ADC molecule Brentuximab Vedotine, developed by Seattle Genetics and Takeda, and recently approved by the FDA and EMEA as Adcetris® (2011, and Nov. 2012, respectively)
- vcPABA has been coupled at its free NH2 to maleimide caproyl for thiol-based conjugation on mAb (cACIO anti-CD30
- vcPABA has been conjugated through its COOH to the Auristatin cytotoxic drug from Seattle Genetics (MMAE) . (see 49)
- the present inventors have used this linker (maleimide caproyl- vcPABA) to conjugate anti-FAP antibodies through thiol-based reaction with the maleimide caproyl, and on the other end, to the cytolysin cytotoxic molecules through its cyclic piperidine with vcPABA (RI or R4 positions of the cytolysin shown in Figure 11) .
- TAM461 (Tubulysin/Cytolysin) : 30.0 mg (0.041 mmol)
- TAM461 and TAM465 were dissolved in anhydrous DMF under dry conditions and the resulting solution was treated with HOBt and DIPEA. The reaction was stirred at RT for 18h. The reaction mixture was concentrated and the resulting oil was purified by column chromatography using 2-6% methanol: DCM to give 35 mg (64%) of
- TAM470 and TAM466 were dissolved in anhydrous DMF under dry
- Rl TAM467, TAM551
- R4 TAM471, TAM553, TAM558
- EG ethylene-glycol spacer
- TPI tubulin polymerization inhibition assay
- CPA cell proliferation arrest on HT1080 cells
- TAM334 In vitro activity of parental cytolysin TAM334 is within the same range of other payloads currently used for the generation of antibody-drug conjugates such as auristatins (MMAE) or maytansinoids (DM1-DM4) .
- MMAE auristatins
- DM1-DM4 maytansinoids
- PLRP chromatography
- the various drugs produced different levels of aggregation.
- a preliminary conjugation with TAM467 also showed high level of aggregation: at DAR 3.27, SEC purity was already only 67% with 16% of free drug (data not shown) .
- Anti-FAP hu36:TAM471 ADC binding to huFAP fusion protein was analysed by ELI SA, and binding to HT1080-FAP cells by FACS ( Figure 15) .
- FACS Fluorescence Activated Cell Sorting
- ADC-471 and ADC-553 showed low and no FAP-specific cytotoxic activity ( ⁇ and ⁇ IC50 range, respectively) with no difference between HT1080-WT and FAP cells nor anti-tumoral effect in vivo
- ADC-558 presented a 1 nM range FAP-specific cytotoxic activity with a specificity ratio of 500 between FAP(+) and FAP(-) HT1080 cells, and a 40% tumor growth inhibition effect at 2.5mg/kg dose in PDX mouse model for pancreas cancer. No weight loss, nor toxic effect was observed for none of the candidates at this dose (not shown) .
- ADC-558 Maximum tolerated dose (MTD) was performed in normal mice and ADC-558 was found to be non-toxic within 2.5 to 25mg/kg dose range with a weekly treatment for 3 weeks. Doses from 20, 10, and 5 mg/kg were then administrated weekly for 4 weeks to a PDX mouse model (Pancl85) with high FAP expression level and stroma expansion to confirm tumor growth inhibition and full regression efficacy of the ADC-558 conjugate .
- MTD maximum tolerated dose
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DK15705778.7T DK3102244T3 (en) | 2014-02-06 | 2015-02-04 | Antibody drug conjugates and immunotoxins |
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US15/116,430 US10137202B2 (en) | 2014-02-06 | 2015-02-04 | Antibody-drug conjugates and immunotoxins |
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Publication number | Priority date | Publication date | Assignee | Title |
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Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE69710911T2 (en) | 1996-05-31 | 2002-09-19 | Health Research Inc | Monoclonal antibodies against endoglin and their use in anti-angiogenesis therapy |
EP0953639A1 (en) | 1998-04-30 | 1999-11-03 | Boehringer Ingelheim International GmbH | FAPalpha-specific antibody with improved producibility |
CA2401252A1 (en) * | 2000-03-17 | 2001-09-20 | Boehringer Ingelheim Pharma Kg | Human and humanized fap-alpha-specific antibodies |
US20020099180A1 (en) | 2000-03-17 | 2002-07-25 | Klaus Pfizenmaier | Human FAP-alpha-specific antibodies |
WO2005108419A1 (en) | 2004-05-07 | 2005-11-17 | Lea-Ann Kirkham | Mutant cholesterol-binding cytolysin proteins |
NZ550934A (en) | 2004-05-19 | 2010-05-28 | Medarex Inc | Chemical linkers and conjugates thereof |
DE102005036542A1 (en) * | 2005-08-03 | 2007-02-08 | Universität Stuttgart | CTL prodrug |
EP1806365A1 (en) * | 2006-01-05 | 2007-07-11 | Boehringer Ingelheim International GmbH | Antibody molecules specific for fibroblast activation protein and immunoconjugates containing them |
NZ599239A (en) * | 2007-03-14 | 2013-10-25 | Endocyte Inc | Binding ligand linked drug delivery conjugates of tubulysins |
EP2532673A3 (en) | 2007-05-10 | 2014-01-08 | R & D Biopharmaceuticals Gmbh | Tubulysine derivatives |
CN102339094A (en) * | 2010-07-21 | 2012-02-01 | 英业达股份有限公司 | Server |
MA34519B1 (en) * | 2010-08-13 | 2013-09-02 | Roche Glycart Ag | ANTI-FAP ANTIBODIES AND METHODS OF USE |
RU2014124984A (en) * | 2011-12-05 | 2016-01-27 | Идженика Биотерапьютикс, Инк. | COMPOUNDS ANTI-MEDICINAL PRODUCT AND RELATED COMPOUNDS, COMPOSITIONS AND METHODS |
CN104662000B (en) * | 2012-05-15 | 2018-08-17 | 索伦托医疗有限公司 | Drug conjugates and its coupling method and purposes |
PL2872157T3 (en) * | 2012-07-12 | 2020-07-13 | Hangzhou Dac Biotech Co., Ltd | Conjugates of cell binding molecules with cytotoxic agents |
CA2940311C (en) | 2014-01-28 | 2022-12-13 | Tube Pharmaceuticals Gmbh | Cytotoxic tubulysin compounds for conjugation |
GB201402006D0 (en) * | 2014-02-06 | 2014-03-26 | Oncomatryx Biopharma S L | Antibody-drug conjugates and immunotoxins |
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2014
- 2014-02-06 GB GB201402006A patent/GB201402006D0/en not_active Ceased
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US20190105406A1 (en) | 2019-04-11 |
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US10864278B2 (en) | 2020-12-15 |
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JP6619743B2 (en) | 2019-12-11 |
BR112016018005A2 (en) | 2018-02-20 |
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WO2015118030A3 (en) | 2015-10-01 |
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